MMP-2、MMP-9对脑缺血再灌注大鼠紧密连接蛋白影响的实验研究  

Experimental Study of MMP-2,MMP-9 Affect on Tight Junction Proteins in Cerebral Ischemia-reperfusion Rats

作　　者：辛浩琳[1] 郭娜 徐忠信[3] 

机构地区：[1]天津市环湖医院神经内科,300211 [2]抚顺市中心医院神经内科 [3]吉林大学中日联谊医院神经内科

出　　处：《医学理论与实践》2015年第7期841-843,共3页The Journal of Medical Theory and Practice

摘　　要：目的:本研究将探讨在再灌注过程中,MMP-2和MMP-9的激活是否参与claudin-5和occludin蛋白的降解。方法:选取再灌注3h这个再灌注时间点进行研究。通过明胶酶谱法,确定MMPs的活性来源;通过Western blot测定claudin-5,occludin在缺血侧、非缺血侧蛋白水平;通过使用MMPs抑制剂GM6001,进一步验证明胶酶(MMP-2和MMP-9)在脑缺血再灌注过程中对claudin-5和occludin蛋白降解的影响。结果:再灌注3h,明胶酶谱法显示MMP-2比MMP-9有较大的增幅。可见,在再灌注3h,MMP-2是最主要的酶源。在缺血侧,可见claudin-5和occludin蛋白降解片段。MMPs抑制剂GM6001使用可以完全抑制缺血侧及非缺血侧MMP-2和MMP-9的活性,并抑制claudin-5和occludin蛋白的降解。结论:MMP-2和MMP-9介导的claudin-5和occludin蛋白降解是与再灌注损伤相关的血脑屏障破坏的一个重要机制。Objective ：In this study ,we are interested in whether the activated MMP‐2 and MMP‐9 are involved in the degradation of claudin‐5 and occludin proteins .Methods ：First ,by gelatin zymography ,we quantified the activities of MMP‐2 and MMP‐9 in the brain of rats subjected to 2h of middle cerebral artery occlusion followed by 3h of reperfu‐sion .Then ,we future investigate the role of MMP‐2 and MMP‐9 activation in the degradation process of claudin‐5 and occludin proteins .Finally ,using MMPs inhibitor GM6001 ,we further study and verify the effect of gelatinases on the degradation of claudin‐5 and occludin proteins after 3h of reperfusion .Results：Gelatin zymography showed greater in‐creases in MMP‐2 than in MMP‐9 .MMP‐2 was the main enzymatic source at 3h reperfusion .claudin‐5 and occludin ex‐pression decreased at 3h in both hemispheres ,with fragments of both proteins seen in the blots at 3h on the ischemic side .MMPs inhibitor GM6001 completely blocked gelatin cleavage by the improved activities of MMP‐2 and MMP‐9 , and prevent the degradation of claudin‐5 and occludin proteins during the early stage of cerebral ischemia reperfusion , respectively .Conclusion：Our study suggests that MMP‐2 and MMP‐9 mediated degradation of the claudin‐5 and occlu‐din proteins may represent an important mechanism for reperfusion‐associated BBB disruption .

关 键 词：MMP-2 MMP-9 CLAUDIN-5 OCCLUDIN 脑缺血再灌注 

分 类 号：R33[医药卫生—人体生理学]